Nephrin in Human Kidney. Nephrin was detected in immersion fixed paraffin-embedded sections of human kidney using 1.7 µg/mL Sheep Anti-Human Nephrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4269) overnight at 4 °C. Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific labeling was localized to podocytes in glomeruli. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Nephrin by Western Blot. Western blot shows lysates of human kidney tissue under reducing and non-reducing conditions. PVDF membrane was probed with 1 µg/mL Sheep Anti-Human Nephrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4269) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band for Nephrin was detected at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Nephrin, also known as renal glomerulus-specific cell adhesion receptor and nephrosis 1 (NPHS1) is a 185-200 kDa type I transmembrane protein belonging to the immunoglobulin (Ig) superfamily. It is expressed on podocytes and is an essential component of the interpodocyte-spanning slit diaphragm complex. Nephrin forms cis-hetero-oligomeric complexes with Neph1, followed by trans-homophilic interaction with Nephrin on opposing cells. Mutations in the Nephrin gene is the pathogenic cause of congenital nephrotic syndrome. Mature human Nephrin contains a 1033 aa extracellular region and a 165 aa cytoplasmic tail. One potential soluble splice form is known where aa’s 1056-1095 are deleted, eliminating the transmembrane region. Over aa 23-1029, human Nephrin shares 84% and 89% aa sequence identity with mouse and canine Nephrin, respectively.
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